Tetracycline resistance heterogeneity in Enterococcus faecium

Bentorcha, F; Cespédès, Gilda de; Horaud, T

doi:10.1128/aac.35.5.808

Cited by 36 publications

(23 citation statements)

References 32 publications

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“…Therefore, tetL itself did not influence the pattern generated with RAPD1 PCR. The occurrence of tetL in all tetracycline-resistant transconjugants confirms the results of earlier and recent studies showing a wide distribution of tetL in E. faecium (1,5). Additionally, tetL seems to be more easily transferable than tetM, which is often found in Tn916-related elements integrated in the chromosome of enterococci (5).…”

Section: Vol 41 2003 Influence Of Conjugative Elements On Typing Mesupporting

confidence: 88%

Influence of Transferable Genetic Determinants on the Outcome of Typing Methods Commonly Used for Enterococcus faecium

et al. 2003

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A variety of methods is used for a molecular typing of Enterococcus spp. and related gram-positive bacteria including macrorestriction analysis using pulsed-field gel electrophoresis (PFGE), ribotyping, rapid amplification of polymorphic DNA (RAPD), and amplified fragment length polymorphism (AFLP). To test the influence of transferable determinants on the outcome of different typing methods commonly used for enterococci, we established a homogenous strain collection of 24 transconjugants resulting from filter matings with antibiotic-resistant Enterococcus faecium. As expected, AFLP, RAPD, and PFGE all identified our model bacteria as strongly related. However, distinct differences in the resolving and discriminatory power of the tested methods could be clearly addressed. In PFGE, 22 of 24 transconjugants possessed less than a three-band difference to the recipient pattern and would be regarded as strongly related. Three different RAPD PCRs were tested; in two reactions, identical patterns for all transconjugants and the recipient were produced. One RAPD PCR produced an identical pattern for 18 transconjugants and the recipient and a clearly different pattern for the remaining 6 transconjugants due to a newly appearing fragment resulting from acquisition of the tetL gene. AFLP clusters all transconjugants into a group of major relatedness. Percent similarities were highly dependent on the method used for calculating the similarity coefficient (curve-based versus band-based similarity coefficient). Fragment patterns of digested plasmids showed the possession of nonidentical plasmids in most transconjugants. PFGE still could be recommended as the method of choice. Nevertheless, the more-modern AFLP approach produces patterns of comparable discriminatory power while possessing some advantages over PFGE (less-time-consuming internal standards). Plasmid fingerprints can be included to subdifferentiate enterococcal isolates possessing identical macrorestriction and PCR typing patterns.The "gold standard" for molecular typing of enterococci and related gram-positive bacteria, such as staphylococci and lactococci is still macrorestriction analysis via pulsed-field gel electrophoresis (PFGE) (13,14). In recent years, some alternative techniques have been successfully applied to the typing of enterococci below the species level. These include amplification-based methods, such as rapid amplification of polymorphic DNA (RAPD) (16, 26) and amplified fragment length polymorphisms (AFLP) (2, 19). These techniques are now applied more and more because they involve less time, comparably low costs, and only standard equipment. Fragments resulting from RAPD PCR typing are randomly amplified and resolved in common agarose gels. AFLP is used for a wide range of organisms with different applications and has been successfully applied to the study of the genetic relatedness of epidemiological unrelated strains of enterococci (33). AFLPresulting fragments are determined in a capillary or gel sequencer allowing detection of single base p...

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Section: Vol 41 2003 Influence Of Conjugative Elements On Typing Mesupporting

confidence: 88%

Influence of Transferable Genetic Determinants on the Outcome of Typing Methods Commonly Used for Enterococcus faecium

et al. 2003

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“…Using probes containing Tet determinants, the presence and the location of special tet genes was screened in a diversity of microorganisms from human sources. More recently, isolates of staphylococci (27,282), E. faecium (20), E. faecalis (46), Peptostreptococcus species (262), Listeria monocytogenes (255), bacteria isolated from the urogenital tract (268), Campylobacter (216), Ureaplasma urealyticum (34), Actinomyces viscosus, Eubacterium lentum, Mobiluncus curtisii and Mobiluncus mulieris (270), and Mycobacterium and Streptomyces species (239) were screened. Using probes, the authors analyzed the presence of different Tet determinants within the different genera.…”

Section: Detection Of Resistancementioning

confidence: 99%

Molecular Detection of Antimicrobial Resistance

2001

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The determination of antimicrobial susceptibility of a clinical isolate, especially with increasing resistance, is often crucial for the optimal antimicrobial therapy of infected patients. Nucleic acid-based assays for the detection of resistance may offer advantages over phenotypic assays. Examples are the detection of the methicillin resistance-encoding mecA gene in staphylococci, rifampin resistance in Mycobacterium tuberculosis, and the spread of resistance determinants across the globe. However, molecular assays for the detection of resistance have a number of limitations. New resistance mechanisms may be missed, and in some cases the number of different genes makes generating an assay too costly to compete with phenotypic assays. In addition, proper quality control for molecular assays poses a problem for many laboratories, and this results in questionable results at best. The development of new molecular techniques, e.g., PCR using molecular beacons and DNA chips, expands the possibilities for monitoring resistance. Although molecular techniques for the detection of antimicrobial resistance clearly are winning a place in routine diagnostics, phenotypic assays are still the method of choice for most resistance determinations. In this review, we describe the applications of molecular techniques for the detection of antimicrobial resistance and the current state of the art

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“…Genes tet(K) and tet(L) have been detected only in gram-positive bacteria (2,3,9,25,27,28,37). The tet(P) determinant, which is constituted of two overlapping genes, tetA(P) and tetB(P), has been found only in Clostridium spp.…”

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confidence: 99%

Presence of the Listeria tetracycline resistance gene tet(S) in Enterococcus faecalis

Charpentier

Gerbaud

Courvalin

1994

Antimicrob Agents Chemother

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Two hundred thirty-eight tetracycline-and minocycline-resistant clinical isolates of Enterococcus and Streptococcus spp. were investigated by dot blot hybridization for the presence of nucleotide sequences related to tet(S) (first detected in Listeria monocytogenes BM4210), tet(K), tet(L), tet(M), tet(O), tet(P), and tet(Q) genes. The tet(S) determinant was found in 22 strains of Enterococcus faecalis, associated with tet(M) in 9 of these isolates and further associated with tet(L) in 3 of these strains. tet (M)

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Tetracycline resistance heterogeneity in Enterococcus faecium

Abstract: The tetracycline (Tet) determinants, which encode resistance either to tetracyclines without minocycline (Tcr)

Cited by 36 publications

References 32 publications

Influence of Transferable Genetic Determinants on the Outcome of Typing Methods Commonly Used for Enterococcus faecium

Influence of Transferable Genetic Determinants on the Outcome of Typing Methods Commonly Used for Enterococcus faecium

Molecular Detection of Antimicrobial Resistance

Presence of the Listeria tetracycline resistance gene tet(S) in Enterococcus faecalis

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